Withaferin A reverses bile duct ligation-induced liver fibrosis by modulating extracellular matrix deposition: Role of LOXL2/Snail1, vimentin, and NFκB signaling.

Abstract

Herein, we studied the effect of Withaferin A (WFA) in reversing bile duct ligation (BDL)-induced liver fibrosis. BDL was performed on C57BL/6J mice and 2 days later, WFA (1 and 3 mg/kg) was administered for 12 days. Estimation of liver enzymes and assays for lipid peroxidation, reduced glutathione, and nitrite levels were performed. Picrosirius red, Masson's trichrome, and H&E staining were performed to study histological changes. WFA proved to be a holistic intervention for the attenuation and reversal of liver fibrosis. Reduction in inflammatory stimulus and oxidative stress restored the levels of stress-related chaperone Hsp70 (p < .001 vs. BDL) in WFA treated groups. We found 3.59-fold (p < .001) and 1.37-fold (p < .01) reduction in the expression of lysyl oxidase like2 (LOXL2) and Snail1, respectively, in WFA-treated animals as compared with BDL animals. These reductions led to 1.9-fold (p < .001) elevation in levels of E-cadherin signifying the reversal of epithelial to mesenchymal transition by WFA. Further, the reduction in LOXL2 levels enhanced the susceptibility of fibrotic scar toward degradation. The picrosirius red and Masson's trichrome staining done on liver tissue sections supported the above results. We, for the first time, report the role of WFA in modulating the expression of LOXL2 and Snail1 in addition to vimentin inhibition and regulation of NFκB signaling for the treatment of liver fibrosis.